HRP20040232A2 - Pesticidal formulations - Google Patents

Abstract

The present invention relates to an active composition for controlling or eradicating Phthiraptera, Siphonaptera and Acarina Pests, typically in domestic animals, comprising a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone. The invention also relates to the use of the active composition in pesticidal formulations, the formulations themselves and to the various applications of those formulations as pesticides, specifically in controlling all species of Phthiraptera, Siphonaptera and Acarina pests, typically in domestic animals. Such applications include the control of such external Phthiraptera, Siphonaptera and Acarina pests in domestic animals including but not limited to sheep, cattle, poultry, pigs, goats, camelids, horses, dogs and cats.

Description

The field of technology

This invention relates to preparations of pesticidal active compounds suitable for use as active agents in pesticide preparations, to the preparations themselves and to various applications of these preparations as pesticides, especially for controlling all types of Phthiraptera, Siphonaptera and Acarina pests. Such applications include the control of external Phthiraptera, Siphonaptera and Acarina pests in domestic animals including, but not limited to, sheep, cattle, poultry, pigs, goats, camels, horses, dogs and cats, as well as domestic and rural applications of such preparations for pest control .

Background of the invention

Historically, the greatest damage to domestic animals and crops has been and continues to be caused by pests such as insects, fungi, nematodes and microbes. Insects are especially a cause for concern, because they are the most numerous of all living organisms and make up approximately 72% of all animal species. Approximately 1% of insects are considered pests in that they attack humans and/or domestic animals, transmit human, animal and plant diseases, destroy crops, buildings and structures and compete for food and other needs. It is estimated that enormous losses occur worldwide due to the presence of insects.

Domestic animals, which include animals of agricultural value such as sheep, cattle, horses, goats, pigs and other ruminants and monogastric animals are almost without exception susceptible to the activities of pests including insects, mites, acarina, siphonaptera, phthiraptera, anoplura and mallophaga. External parasites such as ticks, lice and fleas irritate animals and can cause economic losses in terms of poor fur, wool or sheepskin quality, poor meat/tissue quality, reduced weight gain and even death as a result of the presence of harmful parasites in the animal.

Losses resulting from insects that cause human and animal diseases are also enormous. In fact, insects are considered carriers of more than 250 viruses that are pathogenic to humans and higher animals. The number of human deaths caused by mosquito-borne diseases such as malaria and lymphatic filariasis is enormous. Flies also transmit human and animal diseases such as trachoma, trypanosomiasis and river blindness.

However, of the nearly one million species of arthropods that include lice, ticks, flies and bedbugs, only a small percentage require control measures.

Until now, the primary method for controlling insects and other pests, especially with regard to domestic animals (such as sheep, cattle, goats, horses and pigs) has been the use of synthetic chemical pesticide preparations. It is estimated that there are at least 35,000 formulated pesticide preparations worldwide, with chemicals as active ingredients. Such pesticides include antimicrobials, larvicides, insecticides, animal baths, avicides and disinfectants.

The extensive use of chemical insecticides since the 1940s has resulted in many problems, including widespread insect resistance, the emergence of secondary pests, hazards to human and animal health, as well as adverse effects on fish and birds, environmental pollution, and the increasing economic costs of new insecticides.

Many insect species have developed resistance to specific insecticides to adapt to changes in control practices. There is a growing pool of insect pests that are developing multiple resistance. Resistance genes that have long persistence in insect genomes prevent the successful reuse of insecticides to control populations of insects with resistance genes.

Pesticide/insecticide residues and their consequent many potential human, animal and environmental risks are also considered one of the major problems arising from chemical use, especially those preparations containing active agents including organophosphates or synthetic pyrethroids. With the exception of microbial insecticides, almost all pesticides result in residues of various chemicals and their degradation products or metabolites that can be present in measurable quantities (ppb to ppm) in food, despite food processing. Residues in meat/tissue are also a greater concern when considering the use of insecticides in farm animals.

Potential human risks from the use of such insecticides include acute toxic reactions to the insecticide, such as poisoning, skin and eye irritation, as well as possible long-term effects such as cancer, birth defects, and reproductive disorders. Acute inhalation toxicity as well as skin penetration are also potential risks. Human health hazards can also arise from repeated exposure to a chemical over a limited period of time.

In particular, currently used active substances of synthetic pyrethroids and organophosphates, which are commonly used in insecticidal preparations to control lice and flies, especially on sheep, are toxic not only to animals, but also to humans who apply them. Exposure of farmers or workers handling pesticide concentrates and larger volumes of pesticides diluted for use is cause for concern.

Furthermore, it is possible for the worker to ingest pesticides not only through the mouth, but also through breathing (eg spray stream) and absorption through the skin (accidental spillage). Of particular concern is the use of organophosphates where accidental exposure causes acute and chronic poisoning affecting the nervous system.

Therefore, the control of insects and other pests is required to be directed away from exclusive reliance on insecticides and towards optimizing insect and pest control that is environmentally friendly and economical (integrated pest control). The application of microbial control in which insects are attacked by pathogens such as viruses, bacteria, fungi and protozoa is favored, because such microbial insecticides are highly selective for insect pests and do not leave toxic residues. However, such microbial insecticides also have their own problems, such as difficulty in application, as well as limiting the natural enemy/parasite/disease to a large area. Further, they also lack short duration of action and extreme specificity that limits general applicability.

Recently, in the field of insecticides/pesticides, biological control has been applied, which involves the release of sterilized male insects. Genetic engineering has also recently been applied through the mass introduction of harmful mutations such as chromosomal translocations. However, such procedures are very expensive and strict criteria are required before implementing the release of sterilized males. There are also known chemosterilizing agents that sterilize large parts of the insect pest population, but they are strong carcinogens, which makes their use impossible.

The use of chemical insecticides and pesticides and their ecological and economic viability, the hazardous nature and potency of residues, as well as the increased resistance of insects and pests, together with the highly toxic levels of many chemical insecticides, result in the search for new substances or approaches to control insects and other pests.

Therefore, there is a need for compounds and their combinations that can be used as active agents in pesticides, especially against insects that attack domestic animals or their environment, and that are effective at low application doses, selective in biological action and have low toxicity and high safety level for humans, crops, economic animals, aquatic organisms and birds. Such compounds and combinations must be environmentally acceptable in that they must have a visibly low impact on the environment, as well as be economically profitable on a large scale. Further, insects or other pests must be weakly resistant or non-resistant to such compounds or combinations.

Fermentation product A83543, also known as spinosin, includes a family of related compounds (spinosins) produced by Saccharopolyspora spinosa. These are natural fermentation products with a good safety profile, in contrast to the previously used synthetic organically derived compounds (such as synthetic pyrethroids, organophosphates, organochlorides and carbamates), and have previously shown exceptional insecticidal activity. Therefore, the term "A83543 compounds", which has the same scope as the phrase "spinozine and derivatives and their analogs", includes components consisting of a 5,6,5-tricyclic ring system, fused to a 12-membered macrocyclic lactone, neutral sugar (2N,3N,4N-tri-O-methylrhamnose) and aminosugar (forosamine). The family of natural compounds A83543 includes the genus disclosed in EPO patent application no. 0375316 and which has the following general formula:

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wherein R 1 is H or a group selected from

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and R 2 , R 4 , R 3 , R 5 and R 6 are hydrogen or methyl; or their acid addition salt when R 1 is not hydrogen.

Pokazalo se da porodica spojeva iz fermentacijskog produkta A83543 sadrži pojedinačne spojeve A83543A, A83453B, A83543C, A83453D, A83543E, A83453F, A83543G, A83453H, A83543J, A83453L, A83543M, A83453N, A83543Q, A83453R, A83543S, A83453T, A83453U, A83543V, A83453W, A83453X. Boeck et al. describes spinozines A-H and J and their salts in US Pat. No. 5,362,634, 5,496,932 and 5,571,901, which are incorporated herein by reference. Mynderse et al. describes spinozines L-N, their N-demethyl derivatives and their salts in US patent no. 5,202,242, which is incorporated herein by reference. Turner et al. describe spinozines Q-T, their N-demethyl derivatives and their salts in US Pat. No. 5,591,606, 5,631,155 and 5,767,253, which are also incorporated herein by reference. The spinozines K, O, P, U, V, W, and Y are described in an article by DeAmicis, C.V. et al. in American Chemical Society Symposium Series: Phytochemicals for Pest Control (1997), Chapter 11 "Physical and Biological Properties of Spinosyns: Novel Macrolide Pest-Control Agents from Fermentation" p. 146-154.

Spinosin A (A83543A) was the first spinosin to be isolated and identified from the fermentation solution of Saccharopolyspora spinosa. Subsequent examination of the fermentation solution revealed that the parent strain of S. spinosa produced a number of spinosins (A83543A to J). Compared to spinosin A, for spinosins B to J there are characteristic differences in the substitution patterns of the amino group on forosamine, at certain positions of the ring and on the neutral sugar. S. spinosa strains produce a mixture of spinosins whose primary components are spinosin A (~85%) and spinosin D (~15%). These are the two spinosins currently known to be most active as insecticides.

Similar to spinosins, macrocyclic lactones have also so far shown exceptional insecticidal activity. Macrocyclic lactones have a complex ring structure and include well-known anthelmintic compounds such as avermectins and milbemycins. Avermectins are isolated from the fermentation products of Streptomyces avermitilis, and ivermectin is a compound semi-synthetically produced by modifying avermectin. The basic structure of avermectin is a 16-membered lactone ring to which three main substituent groups are attached: a hexahydrobenzofuran group, a disaccharide group (at C-13) and a spiroketal ring (C-17 to C-28). Doramectin is a new avermectin. Milbemycins are other compounds, which are not avermectins, but can be considered to come within the class of compounds which are macrocyclic lactones. Milbemycins differ structurally from the avermectin group, mainly in the absence of a disaccharide group at C-13. Milbemycin D and milbemycin 5-oxime are two such macrocyclic lactones. Moxidectin is derived from the fermentation product nemadectin and has a methoxime substituent at C-23.

This invention relates to the discovery of a synergistic combination of pesticide compounds, the preparation and application of specific pesticidally active agents based on the synergistic combination and their use in pesticide preparations against Phthiraptera, Siphonaptera and Acarina pests, especially in domestic animals.

Objects of the invention

Accordingly, it is an object of the present invention to provide a pesticidal composition that is systemically effective against Phthiraptera, Siphonaptera and Acarina pests, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone.

It is also an object of the present invention to provide a pesticidal composition that is systemically effective against Phthiraptera, Siphonaptera and Acarina pests in domestic animals including cattle, camels, pigs, dogs, horses, cats, sheep, goats and poultry, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone.

Another object of the present invention is to provide one or more pesticidal compositions that are systemically effective against Phthiraptera, Siphonaptera and Acarina pests, comprising a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone as an active agent, together with at least one acceptable carrier or diluent.

Another object of the present invention is to provide one or more pesticidal compositions that are systemically effective against Phthiraptera, Siphonaptera and Acarina pests in domestic animals including cattle, camels, pigs, horses, dogs, cats, sheep, goats and poultry, containing a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone as an active agent, together with at least one acceptable carrier or diluent.

The subject of this invention is also the provision of a procedure for eliminating and/or controlling pests of Phthiraptera, Siphonaptera and Acarina in domestic animals including cattle, camels, pigs, horses, dogs, cats, sheep, goats and poultry by applying or applying to said animals a pesticidally active combination of compounds alone or together with an acceptable carrier or diluent, so that said pesticidally active combination is systemically delivered to said Phthiraptera, Siphonaptera and Acarina pests.

The subject of this invention is also the provision of a procedure for eliminating and/or controlling pests of Phthiraptera, Siphonaptera and Acarina by applying to said pests a pesticidally active combination of compounds alone or together with an acceptable carrier or diluent, so that such a combination is systematically present in said pests.

The term 'Phthiraptera' or 'Phthiraptera nametnici', as used herein, refers to members of the insect genus Phthiraptera, which are parasitic during one or more stages of their life cycle, including the immature stage (which is defined to include larval (nymph) forms), the adult stage or both stages still include the eggs of Phthiraptera insects.

The term 'Siphonaptera' or 'Siphonaptera pests', as used herein, refers to members of the insect genus Siphonaptera that are parasitic during one or more stages of their life cycle, including the immature stage (which is defined to include larval forms), the adult stage, or both stage still includes the eggs of Siphonaptera insects.

The term 'Acarina' or 'Acarina nametnici', as used herein, refers to members of the arachnoid genus Acarina that are parasitic during one or more stages of their life cycle, including the immature stage (which is defined to include larval (nymph) forms), adults stage or both stages still includes Acarina eggs.

It is further stated that for the purposes of the present application, the term 'spinosyn or an analog or derivative thereof' is defined to include a single spinosyn factor (A83543A-H, J-W or Y), an N-demethyl or other derivative of a single spinosyn factor, or a salt thereof, or a combination thereof, in accordance with the disclosure of the aforementioned references incorporated herein. As noted above, the term "A83543 compound" is used herein to mean an individual spinosine factor, or an analog, derivative or salt thereof, or a combination thereof.

The term 'control or eradication' is used to refer to the reduction of live insects or arachnids (adult or immature forms) or to the reduction of live insect or arachnid eggs. The extent of the reduction depends somewhat on the speed of application, the active substances used, the prevailing weather conditions and other conditions known to those skilled in the art.

The term 'effective amount', also used herein, means an amount sufficient to cause a measurable reduction in the treated insect or arachnid population.

The word 'carrier' is used in the patent application to include mixtures of carriers, i.e. mixtures of more than one substance.

The term "synergistic," as used herein, is defined to mean a combination of components in which the activity of the combination is greater than the sum of the individual activities of each component of the combination.

The term "macrocyclic lactone" as used herein refers to compounds of the milbemycin and avermectin class, including both naturally occurring compounds and their synthetic derivatives, particularly those mentioned herein and in the cited prior art.

The term 'domestic animal', as used herein, includes animals of agricultural value and social animals, including but not limited to cattle, camels, pigs, dogs, cats, sheep, poultry, horses and goats, as well as other ruminants and monogastric animals.

The term "systemically effective", as used herein, means that it has an effect or action only when present within the system of the pest, such as after ingestion or other administration that results in the systemic presence of the active substance in the pest. The term does not denote an effect or action when present within the host system of a domestic animal, it is limited to activity or effectiveness when present within the body of an intruder.

The essence of invention

A first aspect of the present invention provides an active composition for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina, said composition being systemically effective in said pests and comprising a synergistic combination of at least one A83543 compound and at least one compound that is a macrocyclic lactone.

Another aspect of this invention provides a composition for controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said composition including an effective amount of the systemically effective composition of the first aspect of the invention and a carrier, diluent or adjuvant which is physiologically acceptable to the domestic animal.

A third aspect of the present invention provides an externally applied composition for the control or elimination of Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said composition comprising an effective amount of the systemically effective composition of the first aspect of the invention and a carrier that is physiologically acceptable to the domestic animal.

A fourth aspect of the present invention provides a pesticidal bait composition for the control or elimination of Phthiraptera, Siphonaptera and Acarina pests, said composition comprising an effective amount of the systemically effective composition of the first aspect of the invention and an acceptable carrier.

A fifth aspect of the present invention provides an oral composition for the control or elimination of Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said composition comprising an effective amount of the systemically effective composition of the first aspect of the invention and a carrier, diluent or adjuvant which is physiologically acceptable to the domestic animal.

A sixth aspect of the present invention provides a parenteral composition for the control or elimination of Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said composition including an effective amount of the systemically effective composition of the first aspect of the invention and a carrier, diluent or adjuvant which is physiologically acceptable to the domestic animal.

A seventh aspect of this invention provides a method for controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said method includes external application of an effective amount of a systemically effective composition according to the first aspect of the invention, or a preparation according to the second or third aspect of the invention to a localized area of the external surface of said animal .

The eighth aspect of this invention provides a method for controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests, said method includes applying an effective amount of a systemically effective composition according to the first aspect of this invention, or a preparation according to the second or fourth aspect of the invention directly to said pests.

The ninth aspect of this invention provides a method for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina in domestic animals, said method includes oral administration of an effective amount of a systemically effective composition according to the first aspect of the invention, or a preparation according to the second or fifth aspect of the invention to said animals.

The tenth aspect of this invention provides a method for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina in domestic animals, said method includes parenteral administration of an effective amount of a systemically effective composition according to the first aspect of the invention, or a preparation according to the second or sixth aspect of the invention to said animals.

An eleventh aspect of the present invention provides the use of a systemically effective composition of the first aspect of the present invention for the manufacture of a medicament for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina in domestic animals.

A twelfth aspect of the present invention provides the use of a systemically effective composition of the first aspect of the present invention for the manufacture of a bait composition for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina.

Another aspect of the present invention provides an active composition of the first aspect of the present invention or a composition of any of the second to sixth aspects of the present invention for use in controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals.

A further aspect of the present invention provides an active composition of the first aspect of the present invention or a composition of the fourth aspect of the present invention for use in controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests.

This invention relies on the surprising discovery of a synergistic interaction between spinosyn and macrocyclic lactones. While not wishing to be bound by theory, macrocyclic lactones have been observed to have a primary effect on the insect nervous system by activating inhibitory glutamate receptors, while spinosins primarily activate nicotinic acetylcholine receptors in insect neurons, causing neuronal hyperactivity. However, both spinosines and macrocyclic lactones have a secondary effect on gamma aminobutyric acid (GABA)-regulated chloride channels in arthropod neurons, GABA being an inhibitory neurotransmitter. Therefore, it is possible that when combined together, spinosins and macrocyclic lactones have a synergistic effect on the GABA receptor, leading to actions on the insect nervous system that are unrelated to the primary action of either spinosins or macrocyclic lactones.

In particular, in this invention, it has surprisingly been shown that synergism occurs when spinosine and macrocyclic lactone are administered together to animal pests Phthiraptera, Siphonaptera or Acarina in such a way that these chemicals are ingested or otherwise present systemically in the pests. Surprisingly, this effect was not observed in contact-based in vitro tests such as treated paper or transient immersion in treated solutions. While not wishing to be bound by theory, it is assumed that this difference relates to the mechanisms of action and the nature of both chemicals, which have low vapor pressures and do not easily cross the cuticle of arthropods.

Two active agents are used to achieve intracorporeal presence in the intruders. This effect was not observed in tests relying only on contact.

Typically, a first aspect of the present invention provides an active composition for the control or elimination of Phthiraptera, Siphonaptera and Acarina pests, typically in domestic animals, said composition is systemically effective in said pests and is a synergistic combination of spinosine and a macrocyclic lactone compound, wherein spinosine:macrocyclic lactone compounds present in the range from 1000:1 to 1:1000 mass/mass. In particular, the ivermectin:spinozad ratio would be 1:1000 in a formulation that is systemically effective against heartworm (Dirofilaria immitis) or flies.

However, synergism has also been observed to work at lower ratios. Accordingly, typically, in a systemically active composition of the invention, the spinozine compound:macrocyclic lactone compound are present in a ratio of 100:1 to 1:100 wt/wt.

Therefore, typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 100:1 to 1:100 wt/wt.

Also typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 10:1 to 1:10 wt/wt.

Further typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 9:1 to 1:9 wt/wt.

Even more typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 8:1 to 1:8 wt/wt.

Also typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 7:1 to 1:7 wt/wt.

Also typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 6:1 to 1:6 wt/wt.

Even more typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 5:1 to 1:5 wt/wt.

Even more typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 4:1 to 1:4 wt/wt.

Most typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 3:1 to 1:3 wt/wt.

Even more typically, in the systemically effective composition of the invention, the spinozine compound:macrocyclic lactone compound are present in the range of 2:1 to 1:2 wt/wt.

Even more typically, in the systemically effective composition of the invention, the spinozine compound: macrocyclic lactone compound are present in a ratio of 1:1 mass/mass.

One embodiment of the first aspect of the present invention provides a systemically effective composition that is a synergistic combination of spinozade and avermectin.

Typically, the macrocyclic lactone of the first aspect of the invention is selected from the group consisting of ivermectin (22,23-dihydroavermectin B1 described in EP 295117), abamectin, avermectin A1a, avermectin A1b, avermectin A2a, avermectin A2b, avermectin B1a, avermectin B1b, avermectin B2a , and avermectin B2b. Also typically, the macrocyclic lactone of the first aspect of the invention may be selected from the group of compounds related to the naturally occurring avermectins above, but having a group on the 25-substituent other than isopropyl or (S)-sec-butyl groups, as disclosed in European Patent Applications 0214731, 0284176, 0308145, 0317148, 0335541 and 0340832. Also typically, the macrocyclic lactone of the first aspect of the invention may include moxidectin (and derivatives disclosed in EP 259779A), doramectin and its analogs (described in EP 0214731B), selamectin, eprinomectin, including milbemycin. milbemycin oxime, milbemycin D (Antibiotic B41D) and its analogs (described in US 3,950,360) and nemadectins (described in EP 170006A).

More typically, the macrocyclic lactone of the first aspect of the invention is selected from the group consisting of ivermectin, moxidectin, doramectin, selamectin, milbemycin oxime, milbemycin D, eprinomectin and abamectin.

Even more typically, the macrocyclic lactone of the first aspect of the invention is ivermectin.

Even more typically, the active preparation is therefore a synergistic combination of spinozade and ivermectin, which is systemically effective against Phthiraptera, Siphonaptera and Acarina pests.

Typically in the compositions of this invention, the carrier may be non-aqueous or aqueous and the active composition is suspended, dissolved or dispersed in the carrier. Also typically, the active formulation may be mixed with a pharmaceutically or veterinary acceptable diluent or carrier, which is selected with respect to the intended route of administration and in accordance with standard practice. In one embodiment, the carriers or excipients used in the compositions of the third and fourth aspects of the present invention include powder carriers, solvents, emulsifiers, wetting and dispersing agents, and water. In particular, in the bait compositions of the fourth aspect of the present invention, the carriers or excipients include a food source, an attractant, or both, such as blood or other tissue extracts and typically further include one or more of the following agents: yeast, sugar, pheromones, aromas, fragrances, solvents, wetting and dispersing agents.

In another embodiment, the carriers or excipients used in the oral preparations of the fifth aspect of the present invention include tableting excipients such as starch or lactose, encapsulating excipients or carriers, and excipients commonly used in solutions or suspensions, including water , flavoring and coloring agents. The choice of carrier is of course based on compatibility with the active ingredient, including factors such as pH, moisture content and stability. The choice of carrier is also made depending on the method of application of the preparation - for example, whether it will be applied locally to the domestic animal or whether it will instead be applied directly, such as via a bait for pests Phthiraptera, Siphonaptera or Acarina.

One embodiment of any of the second to sixth aspects of the invention provides a composition for controlling or eliminating pests of Phthiraptera, Siphonaptera and Acarina, said composition includes:

a) from 0.1 to 40% by weight of the active composition from the first aspect of this invention, i

b) from 60-99.9% by weight of the corresponding carrier.

Typically each dose of the composition of the invention would contain 30 µg of each of the spinozine compound and the macrocyclic lactone compound, more typically the composition would contain 1 mg - 1 g of each of the spinozine compound and the macrocyclic lactone compound.

The compositions of the third aspect of this invention are typically liquids and may be formulated as concentrates and then diluted prior to use.

The compositions of the fourth aspect of the present invention, which are bait compositions, are typically solids including powdered, granulated or dehydrated forms. It is also typical that such bait preparations can be in the form of liquids or pastes.

It has long been common practice to control external parasites in sheep, cattle and other domestic animals including, but not limited to, goats, pigs and horses, by localized application of a topical preparation containing an active insecticide/parasiticide and a carrier/adjuvant. Accordingly, typically, the composition of the third aspect of the invention is a pour-on composition comprising an effective amount of the active composition of the first aspect of the invention and a topically acceptable carrier.

It is also typical that the composition for topical application may be a spray or bath or solution such as a solution for injection.

The shedding composition of the third aspect of the present invention is typically a liquid and is usually applied to the exterior of the domestic animal as a line or spot, which when ingested by such Phthiraptera, Siphonaptera and Acarina pests, for example after feeding on the treated epidermis of the domestic animal, acts systemically in that to pests thus providing protection to the animal from both immature and adult forms of pests of Phthiraptera, Siphonaptera and Acarina, such as ticks, fleas and lice, and can also act to reduce the number of live eggs of Phthiraptera, Siphonaptera and Acarina.

Although the preparation is applied locally, to a localized area, the active agent migrates over the surface of the animal and covers its entire external surface.

The carrier (also referred to herein as an 'adjuvant') present in such pour-on compositions of the third aspect of the present invention is formulated to achieve good spreading across the skin and/or penetration into the epidermis of the animal. Until now, commercial pouring preparations are suspensions, emulsifying concentrates or solutions and often contain at least one organic solvent. Solvents commonly used as carriers in such pouring compositions include propylene glycol, paraffins, isoparaffins, aromatics, isopropyl myristate (IPM), glycol ethers, alcohols, and n-propyl alcohol.

It is important to note that the aforementioned topical preparations of this invention can be used to control arthropods from the genera Phthiraptera, Siphonaptera and Acarina in several ways. In view of the topical preparations of this invention which only remain on the external surface of the domestic animal, they will be effective against pests such as biting lice and other pests which feed on the epidermis of the animal and thus ingest the systemically effective preparations. Given the topical compositions of this invention, which after topical application at least partially penetrate the epidermis of a domestic animal, penetrating the extracellular fluid and then spreading via the animal's lymphatic system into its bloodstream, these compositions will be effective against blood-sucking pests such as are fleas, ticks and some lice. These pests will be specifically anesthetized if they ingest blood containing a systemically active composition of the present invention. Accordingly, the topical compositions of this invention can also become (by penetrating the epidermis of the animal host) systemically present on the host, as well as extrinsically present on the host.

Another embodiment of the third aspect of the invention provides a pouring composition for controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals, said composition includes:

(a) from 0.1 to 40% by weight of at least one active agent from the first aspect of this invention, and

(b) of 60-99.9% by weight of a suitable carrier selected from the group consisting of TPM/alcohol, OP/IPM/OSU and GTCC/PMP/CAP where

TPM is tripropylene glycol methyl ether;

P is octyl palmitate or 2-ethylhexyl palmitate which is an excellent lubricant, and can also be used as an emollient and solvent;

P sa can be replaced with OS (octyl stearate or 2-ethylhexyl stearate);

PM is an isopropyl myristate that has excellent spreading and softening properties - it can be used as a substitute for IPP or IPL;

PP is isopropyl palmitate;

PL is isopropyl laurate;

MP is PPG 2 myristyl ether propionate, which spreads rapidly and promotes wetting of other substances;

SU is di-2-ethylhexyl succinate and promotes moisturizing and spreading of lipophilic substances on the skin;

CS is isoacetyl stearate which can be used as an emollient, lubricant and spreading agent;

TCC is glyceryl tri caprylate/capronate which is an excellent carrier or excipient for active substances;

AP is a selected blend of branched chain esters that again act as an emollient and spreading agent; the alcohol could be benzyl alcohol, propyl alcohol, diacetone alcohol or other suitable alcohol.

other possible carriers that can be used in the compositions of this invention include organic and inorganic alcohols, including propylene glycol which is a common carrier for ivermectin, ethanol, propyl alcohol, benzyl alcohol, glycols, as well as detergents.

Typically, the compositions of the third aspect of the present invention may be in the form of a powder, cream, suspension, spray, emulsion, foam, paste, aerosol, ointment, salve or gel. More typically, the composition is a solution, and typically soluble in water.

Typically, the compositions of the second, third, fifth or sixth aspects of the present invention can be effectively applied to domestic animals such as sheep, cattle, goats, camels, pigs, dogs, cats, poultry and horses, other ruminants and monogastric animals.

Typically, the bait preparations of the fourth aspect of this invention will be directly applied in a manner different from that on the surface of domestic animals, to obtain direct contact that will result in the intracorporeal presence of the active agents in the three specified genera of pests, as known to those skilled in the art.

Typically, the casting composition of the third aspect of the present invention is applied by casting in one or several lines or in the form of a spot on the dorsal midline (back) or shoulder of the domestic animal. More typically, the casting composition is applied by casting along the animal's back, following the spine. The pouring composition may also be applied to the animal by other conventional methods including passing the impregnated material over at least a small area of the animal, using commercially available applicators, using a syringe, spraying or using a spray channel.

Typically, approximately 0.1-2000 mg of active ingredient/kg of animal body weight is an effective amount for topical application to domestic animals. Typically, about 2-100 mg of the active composition of the first aspect of this invention will be applied to a cow or sheep (per kg of body weight).

Typically, a composition of the present invention, such as a pour-on composition, is formulated such that the active composition is present at a concentration of about 0.1-40% w/v, more typically 0.1-20% w/v, preferably about 0.5 to 5%, depending on the strength of the active substance.

Typically, an active composition or composition of the present invention is formulated for use such that each of compound A83543 and the macrocyclic lactone is present in a concentration range of about 1-500 ppm. More typically, each of the A83543 compound and the macrocyclic lactone is present in a concentration range of about 1-300 ppm. Even more typically, each of the A83543 compound and the macrocyclic lactone is present in a concentration range of about 1-100 ppm. A concentration of 1-500 ppm is most typical for ready-to-use preparations such as diluted baths and sprays. Pour-on compositions of the present invention will typically have an active agent concentration in the range of 1-100 g/L, more typically 5 to 50 g/L, even more typically 10-25 g/L. With respect to the oral preparations, such as tablets, and the parenteral preparations of this invention, the dose typically administered will be from 0.01 to 50 mg/kg of animal body weight, more typically 0.1 to 20 mg/kg. For the bait compositions of this invention, the concentration is typically from 0.05 to 1000 mg/kg, more typically from 1 to 100 mg/kg.

Typically, only a small volume of pour-on preparation is required to be effective against Phthiraptera, Siphonaptera and Acarina pests, on the order of 0.5-80ml per application, with 10-60ml per application being preferred for larger animals such as livestock and 1-20 ml per application for smaller animals such as sheep, dogs and cats.

Typically, the composition of the fifth aspect of the present invention is in the form of a tablet, capsule, bolus, solution, suspension or other elixir. The preparations may also be sustained-release preparations. Such preparations are made in a conventional way, in accordance with standard pharmaceutical and veterinary practice. Typically, capsules, boluses or tablets can be made by mixing the active ingredient with a suitable finely divided diluent or carrier, which additionally contains a disintegrant and/or binder such as talc, starch or lactose.

Also typically, the preparation of the sixth aspect of the present invention is in the form of a sterile aqueous solution or suspension of the active ingredient, with a parenterally acceptable carrier that is water and, if necessary, other excipients such as salt or glucose present.

In the compositions of this invention that have pesticidal activity against Phthiraptera, Siphonaptera and Acarina pests, the active agent is a combination of at least one compound selected from the class of spinosin compounds (including spinosad) and at least one active agent selected from macrocyclic lactones, including ivermectin, abamectin, moxidectin, doramectin, eprinomectin and milbemycin.

The compositions of this invention may conveniently include one or more additional ingredients such as preservatives, spreading agents, adhesion promoters, active solubilizers such as oleic acid or lactic acid, viscosity modifiers, UV blockers or absorbents, coloring agents and stabilizers. such as antioxidants. Suitably, surfactants including anionic, cationic, non-ionic and ampholytic surfactants may also be included in the casting compositions of the present invention.

Isopropyl myristate (IPM), isopropyl palmitate (IPP), caprylic/caproic acid esters and saturated C12-C18 fatty alcohols, oleic acid, oleyl ester, ethyl oleate, triglycerides, silicone oils, and dipropylene glycol mono methyl ether (DPM) are common agents. for spreading used in pouring preparations.

Typically, the methods of the seventh through tenth aspects of the present invention prevent infestations of domestic animals by ticks, lice, fleas, and other Phthiraptera, Siphonaptera, and Acarina pests, including, but not limited to, cattle, sheep, goats, pigs, horses, camels, dogs, cats and poultry and other ruminants and monogastric animals.

Typically, the active compositions, compositions and methods of this invention are effective against immature (including nymphs) and adult forms of Phthiraptera, Siphonaptera and Acarina pests. Intruders do not have to be, although they typically are, present on the host domestic animal.

More typically, the active compositions, compositions and methods of this invention are effective against immature and adult forms of Phthiraptera, Siphonaptera and Acarina pests in domestic animals. Also typically, the active compositions, compositions and methods of this invention are also effective in reducing the number of viable eggs of Phthiraptera, Siphonaptera and Acarina, which may be present in domestic animals.

More typically, a topically, orally or parenterally administered composition of the present invention acts to control body lice (Bovicola Ovis) in sheep, to control similar lice in cattle, goats and camels, to control ticks (eg Boophilus bovis) on cattle and to control Siphonaptera (Ctenocephalides felis and other fleas) in dogs, cats and other domestic animals. Also more typically, topically, orally, or parenterally applied compositions of this invention are effective in controlling lice and ticks in livestock and fleas and in dogs and cats.

The preparations of this invention are made according to known techniques. When the preparation is a solution, the parasite/insecticide is mixed with the carrier or adjuvant, using heating and stirring where necessary. Auxiliary or additional ingredients can be added to the mixture of active substance and carrier or can be mixed with the active substance before adding the carrier.

The compositions of this invention can contain as little as 1 ppm of each macrocyclic lactone compound and spinozine compound per application and still see synergistic action.

Active compositions and preparations from the invention are not toxic to humans and animals, as well as to crops and plants, and the residues in the wool, fibers and tissue of animals that have been treated or to which the preparations have been applied, are at ecologically acceptable levels. Furthermore, the process and preparations of this invention do not cause skin irritation or other toxicity to the end users. Environmental pollution is also minimized.

Also useful, since such spinosin factors and macrocyclic lactones are highly effective at low amounts due to their synergistic action when administered together, this invention is useful against Phthiraptera, Siphonaptera and Acarina pest populations in domestic animals, which have existing levels of resistance to spinosin agents as well. compounds and macrocyclic lactones, when these compounds are used separately.

In general, the application of the preparations of the third aspect of this invention, and the active compositions of this invention is external/local on domestic animals. Such local application can take the form of dipping, showering, spraying, spraying, manual application such as powdering, other application or application of the preparation containing the active substance/active substances. Accordingly, typically, the active compositions of the invention are formulated into a number of topical pesticidal compositions.

Preferably, such topical pesticidal compositions include drips, pour-ons, sprays, baths, powders, lotions, gels, ointments, tars, dressings, towels, creams, sticks, soaps, shampoos, collars, lockets, ear bands and bandages. tail. Casting compositions, including those based on water and organic solvents, as well as emulsions and suspensions, are preferred. As stated above, the preparations may be in a concentrated form that is diluted immediately before use.

Dipping compositions, pouring compositions, fluid spraying compositions, and spraying/spray stream compositions are most preferred.

Wettable powders are another preparation from the invention that is made by mixing the active substance with a powder carrier that is wetted and suspended in water. A surfactant can be added. Wettable powder sprays can be applied to domestic animal housing including poultry houses, barns, pens and piggeries due to their relative safety.

Emulsions are another preparation of the invention, they are solutions of active substances in organic solvents that do not mix with water, usually in a concentration of 1-40%, if necessary with a surfactant to promote emulsification, wetting and spreading. Solvent selection is based on safety for plants, humans and animals, volatility, flammability and cost. Aqueous emulsion sprays from such emulsion concentrates can be used to control Phthiraptera, Siphonaptera and Acarina pests in the household.

As stated above, the administration of the compositions of the fifth aspect of the present invention is via oral administration, while the administration of the compositions of the sixth aspect of the present invention is parenteral, for example intramuscular, subcutaneous or intravenous.

The compositions of the fifth and sixth aspects of this invention will vary with respect to the weight of the active combination depending on the host species of the domestic animal being treated, as well as the body weight. The compositions can be administered at a dosage of from 0.001 mg to about 50 mg per kg of animal body weight, more typically from 0.01 to about 30 mg per kg of animal body weight, and even more typically from 0.1 to about 20 mg per kg of animal body weight. the mass of the animal. It can be given as a single dose or as multiple doses. Higher as well as lower dose ranges are also encompassed by the present invention.

Since the application of the bait preparations of the fourth aspect of the present invention takes place by means other than "on" the domestic animal to obtain direct contact (such as ingestion of the bait by the pest), which will result in the intracorporeal presence of the active agents in the three identified pest genera, as which is known to experts.

The spinozine component of the active preparation of the first aspect of the present invention may be present as a single compound, a mixture of two or more compounds, a mixture including at least one of compounds A83543A and A83543D, or a mixture of at least one A83543 compound together with the dried portion of the fermentation medium in which it was produced.

The macrocyclic lactone compounds used in the present invention include well-known anthelmintic compounds such as avermectins and milbemycins and their derivatives and analogs. As noted above, avermectins are isolated from the fermentation products of Streptomyces avermitilis and the isolation and chemical structure of the eight individual components that make up avermectins (otherwise known as C-076 compounds) is described in detail in British Patent Specification 1573995. Ivermectin is a compound that is semi-synthetically formed chemically by modifying avermectin. Commercially available ivermectin may include, for example, the 25-isopropyl analog of ivermectin. For purposes of the compositions and methods of this invention, avermectins that are lipophilic can be prepared by dissolving the avermectin in an organic solvent such as chloroform, methylene chloride, acetone, and alcohols. The milbemycins, as discussed in detail above, are other compounds that are not avermectins, but which can be considered to belong to the class of compounds that are macrocyclic lactones. Milbemycins differ structurally from the avermectin group, mainly in the absence of a disaccharide group at C-13. Examples of such compounds are described in UK Patent 1390336 and EP Patent Applications 170006, 254583, 334484 and 410615 which are incorporated herein by cross reference.

The spinozine compound may also be present as a salt in the active agent, compositions and methods of this invention. Salts would be prepared using standard salt preparation procedures. For example, spinosin A can be neutralized with an appropriate acid to give an acid addition salt. Acid addition salts of spinosin which can be used in the present invention are useful and include salts formed by reaction with either organic or inorganic acids such as, for example, sulfuric, hydrochloric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, cholic , pamoic, mucinic, glutamic, camphoric, glutaric, glycolic, phthalic, tartaric, formic, lauric, stearic, salicylic, methanesulfonic, benzenesulfonic, sorbic, picric, bemzoic, cinnamic and other similar acids.

In general, emulsifying concentrates of A83543 compounds contain an appropriate concentration of A83543 compound dissolved in an inert carrier that is either a water-miscible solvent or a mixture of an organic, water-immiscible solvent and an emulsifier. The preferred concentration range is 1-500 g/L of the mentioned spinozine compound, even more preferably the concentration range is selected from the following group: 1-400 g/L, 1-350 g/L, 1-300 g/L, 1-250 g/ L, 1-200 g/L, 1-150 g/L, 1-100 g/L, 1-90 g/L, 1-80 g/L, 1-70 g/L, 1-60 g/L , 1-50 g/L, 1-40 g/L, 1-30 g/L, 1-20 g/L, even more preferably 25 g/L. Useful organic solvents include aromatics, including xylenes and petroleum fractions. Other organic solvents can also be used, such as terpene solvents, including rosin derivatives, aliphatic ketones such as cyclohexanone, and complex alcohols such as 2-ethoxyethanol.

Suitable emulsifiers for emulsifiable concentrates can be selected from conventional nonionic surfactants, including ethylene oxide adducts of alkylphenols, and anionic surfactants, including sulfonate alkyl/aryl salts.

Aqueous suspensions (AS) contain suspensions of an active spinozine compound insoluble in water, dispersed in an aqueous carrier in a concentration range of about 1-500 g/L, preferably the concentration range is selected from: about 1-400 g/L, about 1-300 g /L, about 1-250 g/L, about 1-200 g/L about 1-150 g/L, about 1-100 g/L, about 1-50 g/L, about 1-45 g/L, about 1-40 g/L, about 1-30 g/L, preferably about 25 g/L. In general, suspensions are prepared by finely grinding the spinozine compound and mixing it into a carrier consisting of water and surfactants selected from such types as nonionic, sulfonated lignins, and alkyl sulfates. Inert ingredients can also be added.

Aqueous suspensions and emulsions are preferably diluted with water to obtain the desired concentration of spinosin in the final compositions of the invention.

In another preferred formulation, one or more active substances are in the form of a solution of the active substance in water. Sprays are the most common way of applying pesticides on the surfaces of structures such as barns, pens and pig pens. Sprays or dips are the most common method of application on small ruminant species, with water generally as the main carrier.

In the compositions of this invention, particularly in the dip wash compositions, it is preferred that the spinosine compound and the macrocyclic lactone compound are each present at a concentration of about 500 ppm or less. More typically, each is present at a concentration of about 400 ppm or less. Also typically, each is present at a concentration of about 300 ppm or less, more typically 200 ppm or less, even more typically 100 ppm or less, most typically 50 ppm or less.

Best Mode and Other Modes for Carrying Out the Invention

Obtaining the desired compositions of this invention can be achieved by conventional methods, a few examples of which are provided below. The preferred method for obtaining the combination of spinozad and ivermectin of this invention is to either co-formulate the combination or formulate each of the compounds separately, then combine. The compounds may also exist in combination as separate phases. For example, one active compound of the synergistic preparation can be in solution, while the other active compound is in suspension, then such a combination is used to obtain the corresponding preparation from this invention.

EXAMPLES

Introduction

Synergy (or mutual reinforcement) occurs when the combined effect of two pesticides is significantly greater than the additive effect of each one individually in the tested concentrations. For example, the procedure of Sun and Johnson (1960) "Analysis of joint action of insecticides against houseflies", J. Econ. Entomol 53: 887-892, requires the calculation of co-toxicity coefficients. A co-toxicity coefficient of 100 indicates additive action only when 130 or more indicates synergism or enhancement. Alternatively, a generalized linear model approach can be used to generate dose-response lines for each chemical. Lines for each individual chemical are used to predict the efficacy of combinations assuming independent action, ie no synergism or enhancement. The predicted lines are compared with the observed action of the combinations and significant amplification can be detected.

Macrocyclic lactones primarily act on the nervous system of insects by activating inhibitory glutamate receptors, while spinosins primarily activate nicotinic acetylcholine receptors in insect neurons, which causes neuronal hyperactivity. However, both spinosines and macrocyclic lactones have a secondary effect on gamma aminobutyric acid (GABA) which opens chloride channels in arthropod neurons, GABA being an inhibitory neurotransmitter. Therefore, it is possible that when combined, spinosins and macrocyclic lactones have a synergistic effect on the GABA receptor, leading to effects in the insect nervous system that would be unrelated to the primary action of either spinosins or macrocyclic lactones. The aim of these studies was to test the hypothesis on pests of veterinary importance Phthiraptera, Siphonaptera and Acarina.

The following examples illustrate the surprising result that synergism occurs when spinosine and a macrocyclic lactone are co-administered to an animal host in such a way that the chemicals are ingested or present systemically in the host. Surprisingly, this effect was not observed in in vitro assays that rely on contact such as treated paper or transient immersion in treated solutions. It is assumed that this difference is related to the mechanisms of action and the nature of both chemicals, which have a low vapor pressure and do not easily penetrate the armor of arthropods.

Example 1

In-vitro tests to investigate possible synergism between spinozad and ivermectin on lice.

Materials and procedures

Using the procedures of Rugg and Thompson (J Aust Ent Soc, 1993; 32: 1-3), groups of 4 × 15 lice were allowed to feed on ground sheep epidermis containing a range of concentrations of spinosad or ivermectin for 48 hours. . Mortality is tested after 48 hours and LC90 values are determined by probit analysis. Lice are exposed to LC90 concentrations of spinozad or ivermectin and 1/2, 1/4 and 1/8 of the LC90. Additionally, lice are exposed to 1:1, 1:4, 4:1, 9:1, and 1:9 combinations of each chemical as part of the LC90.

For the analysis of treated groups, the method of generalized linear models for predispersed binomial data is used, using the logistic link function. The analysis estimates dose-response curves (on a logarithmic dose scale) for each chemical or combination. Dose-response curves are used to predict the efficacy of combinations, taking into account individual activity and comparing with observed efficacy.

the results

There is significant enhancement in most combinations tested. Enhancement was most pronounced using spinozad:ivermectin ratios of 4:1 and 9:1.

Comparative example

Paper contact test on the ears

Materials and procedures

Groups of 4 × 10-13 lice are exposed to a range of concentrations of spinosad or ivermectin on cotton squares in petri dishes. Mortality is assessed after 18 hours and LC90 values are determined by probit analysis. Lice are exposed to the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 of the LC90. Additionally, lice are exposed to 1:1, 1:4, 4:1, 9:1, and 1:9 combinations of each chemical with each concentration of the chemical that is the LC90 fraction.

the results

All efficacy estimates for group combinations are less than those predicted for single action, i.e. no evidence of synergism.

Discussion

The negative result obtained in the paper contact test compared to the ingestion test supports the assumption that both chemicals must be given to the intruder systemically for reinforcement to occur.

Example 2

In vitro assays to investigate synergism between spinozad and ivermectin in fleas

I) ARTIFICIAL MEMBRANE SYSTEM FOR ADULT FLEAS.

Materials and procedures

The artificial membrane (artificial dog) system developed by Wade and Georgi (J Med Entomol 1988; 25:186-190) allows adult fleas to feed on blood and is suitable for detecting the effect of systemic insecticides. Groups of 25 fleas are allowed to feed on citrated blood containing a range of concentrations of spinosad or ivermectin for 24 hours. Mortality is assessed after 24 hours and LC90 values are determined by probit analysis. Fleas are exposed to the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 of the LC90. Additionally, fleas are exposed to 1:1, 1:4, 4:1, 9:1, and 1:9 combinations of each chemical with each concentration of the chemical that is the LC90 fraction.

the results

There is a significant enhancement in most of the tested combinations. Enhancement is most pronounced with the use of 4:1 and 9:1 ratios of spinozad:ivermectin.

II) FEEDING STUDY OF FLEA LARVAE

Materials and procedures

The procedures of El-Gazzaro et al. were used. (J Med Entomol 1986; 23: 651-654). Groups of 50 flea larvae are allowed to feed on larval rearing medium containing a range of concentrations of spinosad or ivermectin for 4 weeks. After 4 weeks live hatched adults, cocoons and dead pupae are counted. Larval mortality is assessed and LC90 values are determined by probit analysis. Larvae are exposed to the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 LC90. Additionally, larvae are exposed to 1:1, 1:4, 4:1, 9:1 and 1:9 combinations of each chemical with each concentration of the chemical being the LC90 fraction.

the results

There is a significant enhancement in most of the tested combinations.

Example 3

In vitro assays for testing synergism between spinozad and ivermectin on cattle ticks

I) LARVAE SUBMERSION STUDIES.

Materials and procedures

The larval immersion test described by Sabatini, Kemp et al (Vet Parasitol 2001; 95:53-62) was used, except that the larvae were dried for 12 hours first to induce them to absorb the immersion fluid when immersed. Groups of 500 larvae are immersed in liquids containing a range of concentrations of spinozad or ivermectin for 20 minutes, then transferred to pieces of paper. Mortality is assessed after 24 hours and LC90 values are determined by probit analysis. Tick larvae are exposed to the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 LC90. Additionally, larvae are exposed to 1:1, 1:4, 4:1, 9:1 and 1:9 combinations of each chemical with each concentration of the chemical being the LC90 fraction.

the results

There is a significant enhancement in most of the tested combinations. Enhancement is most pronounced with the use of 4:1 and 9:1 ratios of spinozad:ivermectin.

II) STUDY OF INJECTION IN ADULTS.

Materials and procedures

Adult fed ticks are injected with 2 microliters of a range of concentrations of spinosad or ivermectin. Data on egg laying and larval incubation are collected according to standard procedures - for example Sabatini and Kemp (Vet Parasitol 2001 ; 95:53-620) and used to calculate LC90 values. Adults are injected with the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 of the LC90. Additionally, adults are injected with 1:1, 1:4, 4:1, 9:1, and 1:9 combinations of each chemical with each concentration of the chemical that is the LC90 fraction.

the results

There is a significant enhancement in most tested combinations, especially in the 4:1 and 9:1 ratios of spinozad:ivermectin.

Comparative example

Immersion of tick larvae without absorbing the immersion liquid

The larval immersion test described by Sabatini, Kemp et al. (Vet Parasitol 2001; 95:53-62) was used. Groups of 500 larvae are immersed in liquids containing a range of concentrations of spinozad or ivermectin for 20 minutes, then transferred to pieces of paper. Submergence mortality is assessed after 24 hours and LC90 values are determined by probit analysis. Tick larvae are exposed to the LC90 concentration of spinozad or ivermectin and 1/2, 1/4 and 1/8 LC90. Additionally, larvae are exposed to 1:1, 1:4, 4:1, 9:1 and 1:9 combinations of each chemical with each concentration of the chemical being the LC90 fraction.

the results

A dose-response relationship was observed for both compounds. Efficacy estimates for combinations are less than those predicted for single action, i.e. no evidence of potentiation.

Discussion

The negative results for enhancement in the assay in which uptake of the test medium does not occur, compared to the positive evidence of enhancement where the larvae do absorb the test medium, supports the assumption that both chemicals must be present systemically in ticks for enhancement to occur.

Examples of spinosad ivermectin synergistic preparations for controlling pests Phthiraptera, Siphonaptera and Acarina

Example 4

Preparation for pouring

Ingredients g/L

Spinozad 20

Ivermectin 5

Antioxidant, such as BHT 0.5

Crodamol IPM 15

Crodamol OSU 15

Crodamol OP up to 100%

The formulation is applied to the dorsal midline of the animal from the head to the base of the tail using an applicator, usually a self-loading dispensing gun with a nozzle to release a narrow or wide strip of formulation the length of the back. The preparation is applied in an amount of 0.2 mL per kilogram of body weight. Alternatively, a certain volume is applied for each body weight class – eg for sheep 10 ml for animals weighing less than 30 kg, 15 for animals 31-50 kg and 20 ml for animals 51+ kg. Sheep and other fiber-producing animals should be treated within 24 hours of shearing or fiber collection.

Example 5

Suspension concentrate, 20 g/L spinosad, 5 g/L ivermectin

Active chemicals are ground into fine particles using a ball mill.

% wt/wt

Spinozad 2

Ivermectin 0.5

Propylene glycol 10

Surfactant, eg Pluronic P123 2

Mineral thickener, eg Veegum 2

Xanthan gum, eg Rhodopol 23 0.2

Antimicrobial, eg Agent Dowicil 75 0.2

Antifoam agent, eg Antifoam C 0.1

Deionized water up to 100%

The suspension concentrate (SC) is diluted 1:1000 in water and used to fill the bath or sauce. The chemical is applied to the animals by immersion. Alternatively, a shower bath or stream of spray can be used to wet the animal to the skin. Sheep can be treated using a hand-held spray brush to pump diluted chemicals into the wool. For wound dressings, diluted chemicals can be poured into the wound. A certain number of animal species can be treated by spraying with a diluted product, to control Phthiraptera, Siphonaptera and Acarina pests that attack domestic animals.

Example 6

Emulsifying concentrate 20 g/L spinosad, 5 g/L ivermectin

Active chemicals are ground into fine particles using a ball mill.

% wt/wt

Spinozad 2

Ivermectin 0.5

Antioxidant, eg BHT 0.5

10% mixture of ionic and non-ionic surfactants,

for example Toximul 3453F 6.8

Toximul 3454FA 3.2

Aromatic hydrocarbon solvent,

for example Solvesso 150, to get 100%

Emulsifying concentrate (EC) is diluted 1:1000 in water and used to fill a bath or sauce. The chemical is applied to the animals by immersion. Alternatively, a shower bath or stream of spray can be used to wet the animal to the skin. Sheep can be treated using a hand-held spray brush to pump diluted chemicals into the wool. For wound dressings, diluted chemicals can be poured into the wound. A certain number of animal species can be treated by spraying with a diluted product, to control Phthiraptera, Siphonaptera and Acarina pests that attack domestic animals.

Example 7

Oral preparation of tablets for dogs for a dog of 20 kg, 500 mg tablets.

Ingredients g/kg

Core tablets

Spinozad 400

Ivermectin 40

Binder, eg povidone 24

Binding/disintegrating agent eg sodium starch glycolate 20

Lubricant, eg magnesium stearate 7

Lining

Film forming agent hydroxypropyl methylcellulose 25

Plasticizing agent, eg glycerin 4

Coloring agent 20

Example 8

Injection preparation

Ingredients g/L

Spinozad 100

Ivermectin 20

Solvent, eg propylene glycol 350

Solvent, eg glycerol formalin 500

Preservative 0.1

Antioxidant 0.1

Example 9

Bait preparation

Ingredients g/kg

Spinozad 0.1

Ivermectin 0.05

Ground dried beef blood 100

Ground dried dog food 400

Yeast 1

Preservative 0.01

Vermiculite 500

Claims (24)

1. A systemically effective composition for controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests, characterized in that it contains a synergistic combination of at least one A83543 compound and at least one macrocyclic lactone. 2. Active composition according to claim 1, characterized in that the ratio of A83543 compound: macrocyclic lactone is in the range of 1000:1 to 1:1000 mass/mass. 3. Active composition according to claim 1, characterized in that the ratio of A83543 compound: macrocyclic lactone is in the range of 10:1 to 1:10 mass/mass. 4. Active composition according to claim 1, characterized in that said A83543 compound is selected from the group consisting of any spinozine compound and their salts, and a mixture of any two or more spinozine compounds including spinozade. 5. Active composition according to claim 1, indicated by the fact that said macrocyclic lactone is selected from the group consisting of ivermectin, abamectin, avermectin A1a, avermectin A1b, avermectin A2a, avermectin A2b, avermectin B1a, avermectin B1b, avermectin B2a, avermectin B2b, moxidectin , doramectin, selamectin, eprinomectin and milbemycin. 6. Active composition according to claim 1, characterized in that said A83543 compound is spinosad and said macrocyclic lactone is ivermectin. 7. Active composition according to claim 1, indicated by the fact that said pests are present in domestic animals. 8. A preparation for controlling or eliminating Phthiraptera, Siphonaptera and Acarina pests in domestic animals, characterized in that said preparation includes an effective amount of systemically effective composition according to claim 1 and a carrier, diluent or auxiliary agent physiologically acceptable for the domestic animal. 9. The preparation according to claim 8, characterized in that the active composition is present in the preparation in a concentration of about 0.1-40% mass/volume. 10. Preparation according to claim 8, characterized in that the preparation is administered orally to one or more domestic animals. 11. Preparation according to claim 8, characterized in that the preparation is administered parenterally to one or more domestic animals. 12. Preparation according to claim 8, characterized in that said preparation is externally applied to one or more domestic animals. 13. Preparation according to claim 8, indicated by the fact that said preparation is a local preparation for application to domestic animals, said local preparation is selected from the group consisting of preparations for dripping, for pouring, sprays, baths, powders, lotions, gels, ointments, tar ointments, dressings, towels, creams, sticks, soaps, shampoos, collars, lockets, ear bands, dips, liquid jets, squirts/sprays and tail bands. 14. Preparation according to claim 8, characterized in that said preparation is a pouring preparation, which is applied to a localized area of the external surface of the domestic animal. 15. Preparation according to claim 8, characterized in that about 0.1-2000 mg of active agent/kg body weight of the animal is effective for local application on domestic animals. 16. A bait preparation for controlling or removing Phthiraptera, Siphonaptera and Acarina pests, characterized in that said preparation includes an effective amount of a systemically effective composition according to claim 1 and an acceptable carrier, diluent or adjuvant. 17. Method for controlling or preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, characterized in that said method includes external application of an effective amount of the active composition according to claim 1 or preparation according to claim 8 at least on a localized area of the external surface of said animal. 18. Method for controlling or preventing Phthiraptera, Siphonaptera and Acarina pests, characterized in that said method includes application of an effective amount of active composition according to claim 1 or preparation according to claim 8 or claim 16 to said pests. 19. Method for controlling or preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, characterized in that said method includes oral administration of an effective amount of active composition according to claim 1 or preparation according to claim 8 to said animals. 20. Method for controlling or preventing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, characterized in that said method includes parenteral administration of an effective amount of active composition according to claim 1 or preparation according to claim 8 to said animals. 21. The use of an active composition according to claim 1, indicated by the fact that it is used for the production of a drug for preventing or controlling Phthiraptera, Siphonaptera and Acarina pests in domestic animals. 22. The use of the active composition according to claim 1, indicated by the fact that it is used for the production of a bait preparation for preventing or controlling Phthiraptera, Siphonaptera and Acarina pests. 23. Active composition according to claim 1 or preparation according to claim 8, characterized in that it is used to prevent or control Phthiraptera, Siphonaptera and Acarina pests in domestic animals. 24. A preparation for controlling or removing Phthiraptera, Siphonaptera and Acarina pests in domestic animals, characterized by the fact that said preparation contains an effective amount of a systemically effective composition according to claim 1 and a carrier, diluent or auxiliary agent physiologically acceptable for a domestic animal, which is used as a bait , for oral or parenteral administration to domestic animals.